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At least 145 records · Page 8

A novel approach to build algal consortia for sustainable biomass production

In the last decade, microalgae have reemerged as a feedstock for biofuels and a diverse suite of bioproducts. Yet, considerable challenges must be overcome before algal biofuels and bioproducts become technoeconomically viable. At present, single algal strain selected for particular phenotypic traits, such as maximum specific growth rate or lipid content, are commonly scaled for cultivation in open, outdoor raceway ponds due to the low capital costs of these systems. Although this monoculture approach may maximize the production of end products, monocultures are particularly susceptible to crashes associated with environmental and biological variability. An approach that has been proposed to generate more productive and stable microalgal crops is the use of eco-engineered communities, or consortia. Yet, attempts to construct productive consortia have not been consistently successful. We argue that failures stem from the lack of an eco-engineering approach to design species combinations. Here, we used an in silico method to build consortia before testing their performance against monocultures. Focusing on consortia of Nannochloropsis and Microchloropsis, we measured growth of strains along gradients of light, temperature, and salinity and used a functional dispersion approach to generate over 8000 functionally-diverse consortia combinations. We tested the 50 most functionally diverse consortia in a laboratory experiment and found that consortia overwhelmingly outperformed monocultures. Indeed, overyielding (OY) and a positive net biodiversity effect (NBE) was found respetively in 8%-86% and 88-92% of consortia combinations over the different experimental phases. To our knowledge, this is the first application of an in silico approach to design functionally diverse consortia before laboratory and field testing. Furthermore, our results highlight the importance of employing a functional diversity approach for consortia design.

09 BIOMASS FUELS↗

Oxygen stress mitigation for microalgal biomass productivity improvement in outdoor raceway ponds

Dissolved oxygen (DO) concentrations at supersaturated levels are commonly observed in outdoor microalgae cultures due to photosynthesis during periods of rapid growth. Paradoxically, the photosynthetically generated oxygen is inhibitory and lowers biomass productivity, therefore, increasing the cost of microalgal biofuels. In this study, photosynthetic rates of Monoraphidium minutum 26B-AM and Scenedesmus obliquus UTEX393, two of the top DISCOVR strains, were compared at three DO levels, 0, 8, and 30 mg L -1 . Increasing DO from 0 to 30 mg L -1 caused a 36.6% and 26.1% reduction in photosynthetic rate for M. minutum 26B-AM and S. obliquus UTEX393, respectively. To improve the outdoor pond biomass productivity of each strain, three oxygen stress mitigation methods were tested. The first method raised the dissolved bicarbonate concentration by an order of magnitude to favor bicarbonate transport into the cell, increasing the CO 2 :O 2 ratio near RuBisCO, thus reducing photorespiration. Although this method increased the observed photosynthesis rate of M. minutum 26B-AM by 26% and S. obliquus UTEX393 by 13% in laboratory testing, it failed to improve biomass productivity under outdoor pond conditions for both strains. The second method added sodium sulfite as an oxygen scavenger to chemically lower the DO concentration in the culture. Chemical reduction of dissolved molecular oxygen did not significantly improve the biomass productivity of M. minutum 26B-AM; however, a 10% improvement in biomass productivity for S. obliquus UTEX393 was observed. The final method sparged cultures with air to strip oxygen out of the liquid medium. This did not improve the biomass productivity of M. minutum 26B-AM, likely due to unfavorable weather conditions during cold season cultivation; however, an improvement of 36% in biomass productivity was achieved for S. obliquus UTEX393 during the warm season cultivation. Of the three tested methods, air-sparging was the most effective and practical for improving outdoor biomass production.

59 BASIC BIOLOGICAL SCIENCES↗

Axenisation of oleaginous microalgal cultures via anoxic photosensitisation

Growing interest in sustainable food and biofuel research has necessitated high quality axenic oleaginous microalgal strains. Unfortunately, most strains available in culture banks contain commensal microbes such as bacteria and the default decontamination method involves antibiotic treatment which has begun to exacerbate the emergence of antibiotic resistance. To overcome this problem, anoxic photosensitisation was investigated as an alternate approach. Four oleaginous microalgal species (Tetradesmus obliquus, Desmodesmus armatus, Chlorella vulgaris and Nannochloropsis limnetica) were incubated in varying concentrations of Rose Bengal (0 μM, 1 μM, 3 μM or 9 μM) either in normal (oxic) or anoxic conditions, for 72 h under light (8.85 ± 0.4 W/m 2 ) in a specially designed heterotrophic growth complex (HGC) medium, followed by 72 h in standard Bold's Basal Medium (BBM). Commonly used antibiotics-based protocol was used as the control method. Post treatment, cell numbers and percentage populations were counted with Flow Cytometry, and viability was tested using standard plating methods using BBM and LB. Additionally, the contaminating microbes in the cultures were profiled using 16Ss rRNA sequencing. Anoxic conditions were able to significantly decrease bacterial content, albeit with an equally detrimental effect on the microalgal population. Although the responses differed between the microalgae, anoxic incubation along with Rose Bengal at 3 μM was able to completely decontaminate N. limnetica and C. vulgaris, while D. armatus and T. obliquus could be decontaminated with an additional streak-plating step. None of the cultures could be decontaminated using antibiotics treatment. These results suggest that axenisation of microalgal cultures was largely due to anoxy, that was synergistically enhanced by Rose Bengal at a concentration of ≥3 μM.

59 BASIC BIOLOGICAL SCIENCES↗

PERCEPTIVE: an R shiny $\underline{p}$ipelin$\underline{e}$ for the p$\underline{r}$edi$\underline{c}$tion of $\underline{ep}$igenetic modula$\underline{t}$ors $\underline{i}$n no$\underline{v}$el sp$\underline{e}$cies

Epigenetic processes are central to regulating gene expression, genome stability, and metabolic function across the tree of life; yet, their roles remain underexplored in microalgae, especially as new species continue to be identified and characterized. This is likely due to the cumbersome nature and species-dependent attributes of epigenetic wet-lab methodologies, which preclude the rapid identification of epigenetic modifications and modulators. However, there is high conservation of epigenetic processes from budding yeast to humans; in many cases, one may infer how behavior and function are epigenetically regulated in novel species by identifying epigenetic modulators, or the proteins responsible for conferring epigenetic modifications. Here, to this end, we have developed a graphical software package, titled PERCEPTIVE (pipeline for the prediction of epigenetic modulators in novel species). This platform solely uses the genomic sequence of an algal species, and preexisting information from other model organisms, to predict the epigenetic modulators and associated modifications in algae. Predictions are presented to the user in a graphical interface, which provides literature-based interpretation of results, enabling users to quickly understand potential epigenetic processes in their algal species of interest and plan follow-up experiments. To test PERCEPTIVE, we predicted epigenetic modulators in several feedstock candidate algae species. To validate these predictions, wet-lab studies were performed, including mass spectrometry; these results underscore the high accuracy of PERCEPTIVE predictions. Overall, PERCEPTIVE represents a powerful in silico tool for the research and manipulation of algal species, which does not require a priori knowledge of epigenetics and is accessible to a broad set of investigators.

59 BASIC BIOLOGICAL SCIENCES↗

Microalgal lipids: biochemistry and biotechnology

Microalgae, including cyanobacteria, are a genetically diverse and biochemically diverse group of mostly photosynthetic organisms that can be found in nearly every ecosystem. They produce a broad range of compounds, including structural and bioactive lipids with various roles in the metabolism of the cell and potential applications in biotechnology. Here, the majority of lipids are synthesized in the chloroplast using acetate to produce fatty acids and terpenoids via the acetate pathway and methylerythritol phosphate deoxy-xylulose phosphate pathway, respectively. This review will highlight the major groups of microalgal lipids as well as their applications in food, fuels, and specialty chemicals. Future directions include the development of microalgal chassis organisms and molecular tools to manipulate lipid synthesis for the enhanced production of target metabolites.

cyanobacteria↗

Morphotype-resolved characterization of microalgal communities in a nutrient recovery process with ARTiMiS flow imaging microscopy

Microalgae-driven nutrient recovery represents a promising technology for phosphorus removal from wastewater while simultaneously generating biomass that can be valorized to offset treatment costs. As full-scale processes come online, system parameters including biomass composition must be carefully monitored to optimize performance and prevent culture crashes. In this study, flow imaging microscopy (FIM) was leveraged to characterize microalgal community composition in near real-time at a full-scale municipal wastewater treatment plant (WWTP) in Wisconsin, USA, and population and morphotype dynamics were examined to identify relationships between water chemistry, biomass composition, and system performance. Two FIM technologies, FlowCam and ARTiMiS, were evaluated as monitoring tools. ARTiMiS provided a more accurate estimate of total system biomass, and estimates derived from particle area as a proxy for biovolume yielded better approximations than particle counts. Deep learning classification models trained on annotated image libraries demonstrated equivalent performance between FlowCam and ARTiMiS, and convolutional neural network (CNN) classifiers proved significantly more accurate when compared to feature table-based dense neural network (DNN) models. Across a two-year study period, Scenedesmus spp. appeared most important for phosphorus removal, and were negatively impacted by elevated temperatures and increase in nitrite/nitrate concentrations. Chlorella and Monoraphidium also played an important role in phosphorus removal. For both Scenedesmus and Chlorella, smaller morphological types were more often associated with better system performance, whereas larger morphotypes likely associated with stress response(s) correlated with poor phosphorus recovery rates. Furthermore, these results demonstrate the potential of FIM as a critical technology for high-resolution characterization of industrial microalgal processes.

59 BASIC BIOLOGICAL SCIENCES↗

Applied genomics for industrially relevant microalgal strain development & optimization: characterizing genotype-phenotype relationships towards scalable environment-enhancing energy systems

Multiple planetary boundaries considered a safe operating space for humanity have been exceeded in recent years, with twice as many boundaries transgressed in 2023 as in 2009. Bioenergy systems are unique in that they can interact with and improve many of the transgressed boundaries directly, including multiple geochemical cycles, water and land use, and climate change. Among bioenergy systems, microalgae-based environment enhancing energy (E 2 -energy) are promising bioenergy systems for drop-in biofuels, valuable materials, chemicals and therapeutics, while making deep emission cuts and remediating wastewater, all without competing for agricultural resources.

09 BIOMASS FUELS↗

The phycosphere and its role in algal biofuel production

Oleaginous microalgae have become a focus for large-scale biofuel production due to their ability to accumulate large quantities of lipids. However, production is currently limited by cost and predation. At present, algal biofuel cultivation is optimized through starvation, supplementing media with nutrients, or genetic engineering; these methods can often be costly with little to no increase in lipid production or the culture’s defense. Investigating the phycosphere of algal-bacterial interactions may overcome these current barriers to large-scale production. The phycosphere of algal-bacterial interactions have formed over millions of years through mutualistic and symbiotic relationships and can provide a more direct source of nutrients compared to adding the nutrients in bulk. The most promising of these interactions include the production of phytohormones and quorum signaling compounds that alter the behaviors of the consortia. Phytohormones can improve algal growth rates, lipid production, and stress resistance. Quorum signaling could create consortia capable of warding off invaders—such as rotifers—while self-regulating and altering behavior based on population density. Mechanisms within the algal phycosphere present many opportunities for the development of novel engineering strategies to further improve algal lipid production and operational costs. This review outlines previous preliminary phycosphere research as well as posing possible opportunities to be pursued in future biofuel production.

09 BIOMASS FUELS↗

Potential of U.S. Marine and Offshore Wind Energy to Power Marine Carbon Dioxide Removal

The goal of this project was to provide a preliminary feasibility assessment of powering different marine carbon dioxide removal (mCDR), carbon capture (mCC), and carbon sequestration (mCS) strategies with marine energy. The investigation focused on methods with clear power requirements at sea beyond monitoring such as artificial upwelling, deep ocean storage, electrochemical (eChem) mCDR and mCC, offshore microalgae cultivation, and seaweed farming and sinking. The project found that eChem mCDR powered by marine energy and offshore wind energy available in the United States could meet global CDR scales needed by 2040 and 2050 to limit warming to 1.5 degrees C by 2100 and marine energy alone could greatly contribute to reaching scales needed by 2040. Note that this preliminary estimate assumes that it is possible to harvest all the marine and offshore wind resources available in the United States with existing technology options. Though the biological methods were limited in scale they still hold promise in developing carbon negative fuels and products which can reduce emissions in the short term.

afforestation↗

An Algal Nutrient‐Replete, Optimized Medium for Fast Growth and High Triacylglycerol Accumulation

Microalgae are promising sources to sustainably meet the global needs for energy and products. Algae grow under different trophic conditions, where nutritional status regulates biosynthetic pathways, energy production, and growth. The green alga Chromochloris zofingiensis has strong economic potential because it co-produces biofuel precursors and the high-value antioxidant astaxanthin while accumulating biomass when grown mixotrophically. As an emerging reference alga for photosynthesis, metabolism, and bioproduction, C. zofingiensis needs a defined, optimized medium to standardize experiments during fast growth for batch cultivation. Because the interplay of glucose treatment (+Glc) and mineral deficiency influences photosynthesis, growth, and the production of lipids and astaxanthin, we designed a replete nutrient medium tailored to the C. zofingiensis cellular ionome. We combined inductively coupled plasma mass spectrometry (ICP-MS) and +Glc growth curves to determine a medium that is nutrient replete for at least 5 days of +Glc logarithmic growth. We found that there are high nutritional needs for phosphorus and sulfur during mixotrophy. Iron was the only element measured for which the cellular concentration correlated with exogenous concentration and was iteratively adjusted until the cellular ionome was consistent through the logarithmic growth phase. This Chromochloris-Optimized Ratio of Elements (CORE) medium supports fast growth and high biomass and lipid accumulation without causing excess nutrient toxicity. This defined, nutrient-replete standard is important for future C. zofingiensis investigations and can be adapted for other species to support high biomass for batch cultivation. The method used to develop CORE medium shows how ionomics informs replicable media design and may be applied in industrial settings to inform cost-effective biofuel production.

Jeffers, Tim L. [University of California, Berkele↗

Cultivating clarity: understanding the impact of land cost assumptions on biofuel viability

The transition to sustainable energy has increased interest in biofuel production to reduce greenhouse gas emissions, decrease reliance on imported oil, and ensure energy resilience. Here, this study examines the often-overlooked impact of land cost assumptions on the economic viability of biofuel production. Using a discounted cash flow techno-economic framework, we evaluated three land cost scenarios—no land costs, land rental costs, and land purchase costs—across six bioenergy feedstocks (corn, soybeans, switchgrass, miscanthus, poplar, and microalgae) and three biofuel products (corn ethanol, soybean biodiesel, and sustainable aviation fuel) at the county level for the contiguous United States. The analysis reveals substantial variation in minimum fuel selling prices due to these scenarios. High-yield crops like algae showed low sensitivity to land costs, while low-yield crops such as soybeans were highly sensitive. Geographical differences were significant, with minimum fuel selling price increases most pronounced in high-value land regions like the Corn Belt. Case studies further illustrate the influence of local productivity and land costs on economic outcomes across the United States. These findings emphasize the importance of maintaining consistent land cost assumptions in biofuel economic assessments. By quantifying the interplay between land value, crop productivity, and economic feasibility, this study provides essential insights for policymakers and stakeholders to advance sustainable energy solutions.

09 BIOMASS FUELS↗

A selection approach using high‑oxygen and pond-mimicking culture conditions increases biomass productivity in the industrially relevant diatom Nitzschia inconspicua str. hildebrandi

To meet the energy and food challenges of the 21st century, renewable and sustainable sources of fuel and nutrients are required. Microalgae offer promise in meeting these challenges but must achieve higher rates of areal productivity to compete with fully scaled, existing industries. In this study, we used selective O 2 pressure to attain a ~90% increase in areal biomass productivity relative to the parental strain in the diatom Nitzschia inconspicua str. hildebrandi under pond-mimicking conditions with high O 2 stress. The resulting strain (GAI-337) was tested further for dilution time, culture density, CO 2 supplementation, pH, temperature, and dissolved O 2 concentration under outdoor pond-mimicking conditions to improve areal productivities. These experiments yielded an optimum harvest and dilution time just after sunset, ~0.45 g AFDW L -1 initial culture density for maximal productivities, no requirement for CO 2 gas supplementation or pH control, maximal performance under a diel temperature curve going from 24 °C at night to 36 °C during the day, and benefits from some O 2 removal from the culture by bubbling with air. Using pond-mimicking laboratory bioreactors, N. inconspicua GAI-337 achieved ~42 g AFDW m -2 d -1 , placing it among the most productive microalgal strains tested to date. Nutrient limitation experiments resulted in a biomass composition with 36% of AFDW comprised of lipids (measured as FAMEs) that equated to ~160 Gallons of Gasoline Equivalent energy per ton AFDW, highlighting the potential of GAI337 as a promising renewable fuel feedstock strain.

09 BIOMASS FUELS↗

Description of a novel extremophile green algae, Chlamydomonas pacifica , and its potential as a biotechnology host

We present the comprehensive characterization of a newly identified microalga, Chlamydomonas pacifica , originally isolated from a soil sample in San Diego, CA, USA. This species showcases remarkable biological versatility, including a broad pH range tolerance (6–11.5), high thermal tolerance (up to 42 °C), and salinity resilience (up to 2 % NaCl). Its amenability to genetic manipulation and sexual reproduction via mating, particularly between the two opposing strains CC-5697 & CC-5699, now publicly available through the Chlamydomonas Resource Center, underscores its potential as a biotechnological chassis. The biological assessment of C. pacifica revealed versatile metabolic capabilities, including diverse nitrogen assimilation capability, motility and phototaxis. Genomic and transcriptomic analyses identified 17,829 genes within a 121 Mb genome, featuring a GC content of 61 %. The codon usage of C. pacifica closely mirrors that of C. reinhardtii , indicating a conserved genetic architecture that supports a trend in codon preference with minor variations. Phylogenetic analyses position C. pacifica within the core-Reinhardtinia clade yet distinct from known Volvocales species. The lipidomic data revealed an abundance of triacylglycerols (TAGs), promising for biofuel applications and lipids for health-related benefits. Our investigation lays the groundwork for exploiting C. pacifica in biotechnological applications, from biofuel generation to synthesizing biodegradable plastics, positioning it as a versatile host for future bioengineering endeavors.

Alkali tolerant↗

Hydrocyclone pre-processing of wastewater algae: A strategy for inorganic ash separation

Microalgae cultivation on wastewater can provide remediation and generate valuable feedstocks for biofuel production. Wastewater algae typically have a high percentage of inorganic ash, which can reduce yield and quality of biocrude produced during hydrothermal liquefaction (HTL). Here, in this work, we evaluated the ability of hydrocyclone pre-processing to remove inorganic ash from wastewater algae. The pH of the algae slurry was adjusted to 9.5 to encourage the formation of precipitates and create a density differential between ash particles and algal cells. Hydrocyclone processing successfully concentrated ash particles in the underflow fraction and reduced the total ash percentage in the overflow fraction. Overall, hydrocyclone processing reduced the total ash by 21%, while only 8% of organics were lost. Elemental and mineral analysis showed that Mg and P were concentrated in the underflow in the form of baricite (an isomorph of vivianite). Future research should focus on improving vivianite and/or baricite formation, and therefore ash removal, by providing a reducing environment. The addition of multiple hydrocyclones in series could also improve the removal of ash. We concluded that hydrocyclone treatment of wastewater algae is a feasible method to remove inorganic ash, but further process optimization is required.

09 - BIOMASS FUELS↗

Computational modeling of microalgal biofilm growth in heterogeneous rotating algal biofilm reactors (RABRs) for wastewater treatment

Rotating algal biofilm reactors (RABRs) are innovative systems designed to cultivate microalgae biofilms efficiently. In this paper, we have developed a novel mathematical model to accurately capture the growth dynamics of algae biofilms within RABR. By considering the spatial heterogeneity of the RABR, we introduce a PDE-based model that addresses the spatial variations across the substratum, enabling a more accurate simulation of biofilm growth in RABRs. The photosynthesis process is modeled through reactive kinetics, driving the growth of the algae biofilm. To analyze the system's behavior, we employ finite difference numerical methods to solve the complex PDE model. We then conduct extensive numerical simulations to understand algae biofilm growth in the RABR environment under various operational factors and environmental conditions. One primary focus in these simulations is to investigate the impact of various harvesting strategies, harvesting frequencies, light intensity, and light exposure on the overall biomass productivity of the algae biofilm. The numerical results provide valuable insights into optimizing algae biofilm growth and designing harvesting techniques in RABR systems. Our proposed novel mathematical model provides an effective platform for the theoretical investigation and design of RABRs for wastewater treatment.

09 BIOMASS FUELS↗